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翌圣生物科技（上海）股份有限

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供應簡介

Collagenase （from clostridium histolyticum） is a protease with a specificity for the bond between a neutral amino acid （x） and glycine in the sequence Pro-X-Glyc-Pro. This sequence is found in high

詳細介紹

Background（背景）

frequency in collagen and is unique among proteases in its ability to degrade the triplehelical native collagen fibrils commonly found in connective tissue.

The collagenase most commonly used for tissue dissociation is a crude preparation containing clostripiopeptidase A and a number of other proteases, polysaccharidases and lipases. This crude

enzyme is ideally suited for tissue dissociation since it contains the enzyme required to attack native collagen and reticular fibers, in addition to the enzymes which hydrolyze the other proteins, polysaccharides and lipids in the extracellular matrix of connective and epithelial tissues.

Crude collagenase does exhibit lot-to-lot variability and may produce occasional toxicity. Invitrogen

attempted to minimize these difficulties by tissue-typing their crude collagenase lots based upon a correlation between various enzyme levels in each preparation and effectiveness in dissociating certain tissues.

Storage（儲存）: 2 to 8°C （-5 to -20°C after reconstitution）

Avoid moisture and exposure to light.（避免潮濕，暴露在光線下）

Avoid inhalation and skin contact.（避免吸入和皮膚接觸）

This product is intended for cell or tissue disaggregation only.（本產品僅用于細胞或組織分解）

Specifications（產品規格）

Potency（效力）:One unit liberates 1 μM of L-leucine equivalents from collagen in 5 hours at +37°C, pH 7.5.

Types（類型）:Particular enzymatic activities of crude collagenases have correlated with the tissues from which the cells were obtained （or with the uses to which the cells are put）, and as a result of the correlation’s, several formal types have been established.

Type III Selected because of low proteolytic activity. （casein as substrate）

Type IV Selected because of low tryptic activity. （BAEE as substrate）

These selected types have been found to give better performance in preparation of cells from the various tissues as tabulated below. It should be noted, however, that while the results have been greatly improved following this classification, there is still some lot-to-lot variation, and efforts continue in attempting to gain even better control over crude collagenase.

TYPE TISSUES OR CELLS

I Fat cells, Adrenal, Liver

II Heart, Bone, Thyroid, Cartilage, Liver

III Mammary

IV Islet （insulin receptor sites）

Inhibitors: Metal chelating agents such as cysteine, EDTA or o-phenanthroline but not DFP. It is also inhibited by a2-macroglobulin, a large plasma glycoprotein.

Instructions for use（使用說明）

A. Preparing stock and working solution

Dissolve the non-sterile, lyophilized enzyme in HBSS （Cat. No. 14025）. Filter sterilize the solution with a cell culture approved filtration unit. Crude collagenase is most often used in concentrations from 0.1 to 0.5% （W/V） or 50 to 100 U/mL. Once reconstituted use immediay or store frozen. Thaw in refrigerator immediay prior to use.

B. Dissociation of tissue

• Tissue is minced with a sterile scalpel or scissors.

• Wash the tissue several times in HBSS.

• The tissue fragments are soaked at +37°C. Increased efficiency is obtained using a rocker platform and supplementing the digest with 3 mM CaCl2.

C. Organ perfusion

• Digest is prewarmed to +37°C and perfused at a rate preoptimized for the particular organ. Addition of 3 mM CaCl2 increases the efficiency of dissociation.

• Dispersed cells and tissue fragments are separated from larger pieces by passing the mixture through a sterile stainless steel or nylon mesh. Fresh collagenase solution can be added to the fragments if further disaggregation is required.

• Wash several times to eliminate debris and enzyme solution. A density separation step （Nycodenz） will give a cleaner suspension.

• Resuspend the pellet in the culture medium and incubate under predetermined conditions.

Ordering Information（訂購信息）：

貨號	名稱	產地	規格	報價	*（元）	庫存
17100017	Collagenase I 膠原酶 I	Invitrogen	100mg	380	280	是
17100015	Collagenase II 膠原酶 II	Invitrogen	100mg	380	280	是
17104019	Collagenase IV 膠原酶 IV	Invitrogen	100mg	380	280	是
02195109.1	Collagenase I 膠原酶 I	Mpbio	100mg	691.6	280	是
02100502.1	Collagenase II 膠原酶 II	MPbio	100mg	873.6	280	是
	Collagenase IV 膠原酶 IV	MPbio	100mg	768.6	280	是
C9263	Collagenase V 膠原酶 V	Sigma	100mg	607	488	是

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